Figure 6.

NAM supplementation partially rescues GLUT4 expression and fully supports adiponectin expression. 3T3-L1 adipocytes were induced for differentiation and pulsed with Hi glucose, Lo glucose, or Pyr without or with 15 mm NAM and cultured in Lo glucose medium during the P and M phases of their cell culture A, day 3 adipocytes were collected and lysed accordingly, and the supernatants were analyzed via ELISA for levels of total NAD. B, protein from whole-cell lysates of day 9 adipocytes were separated via SDS-PAGE and immunoblotted for GLUT4, adiponectin, and tubulin. C–E, mRNA isolated from day 9 adipocytes were analyzed via qRT-PCR for the relative expression of glut4, pparg, and ap2. 95% confidence intervals were used to determine statistical differences in immunoblot densitometry. One-way analysis of variance was used to identify significant differences in expression and ELISA data. *, differences compared with Hi glucose cells; #, differences compared with Lo glucose cells. ‡, difference comparing NAM treatment with the untreated control.